A peptide-based point-of-care vertical flow assay for the rapid diagnosis of Lyme disease

基于肽的即时垂直流检测用于快速诊断莱姆病

• 批准号: 10404209
• 负责人: Paul Michael Arnaboldi
• 金额: $ 100万
• 依托单位: BIOPEPTIDES, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-24 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10404209
• 关键词: Address; Antibiotics; Antibodies; Antigen Targeting; Antigens; Bacterial Proteins; Binding; Binding Proteins; Biological Assay; Borrelia burgdorferi; Borrelia miyamotoi; Cells; Cellular Phone; Centers for Disease Control and Prevention (U.S.); Clinical; Clinical Treatment; Coupled; DBL Oncoprotein; Data; Defect; Detection; Development; Diagnosis; Diagnostic; Disease; Disease Progression; Early Diagnosis; Early treatment; Epitopes; Family; Generations; Goals; ITGB3 gene; ImProv; Immunoglobulin G; Immunoglobulin M; Individual; Infection; Intervention; Laboratories; Laboratory Diagnosis; Legal patent; Lyme Disease; Machine Learning; Maps; Measures; Membrane; Methods; Microbe; Musculoskeletal System; Nature; Nervous system structure; OspC protein; Paper; Patient-Focused Outcomes; Patients; Peptides; Phase; Physicians; Proteins; Reader; Reporting; Research; Sampling; Serology; Signal Transduction; Small Business Innovation Research Grant; Specificity; Speed; Spottings; System; Technology; Testing; Time; Work; antibody detection; base; biological systems; blind; clinical Diagnosis; cost; cost effective; cross reactivity; decorin binding protein B; design; detection assay; detection platform; diagnostic assay; efficacy testing; improved; innovation; member; multiplex detection; novel strategies; point of care; point of care testing; point-of-care detection; point-of-care diagnostics; portability; preservation; prevent; prototype; public health relevance; rapid diagnosis; rapid test; relapsing fever borrelia; screening

Abstract Project Summary There is an obvious need for new approaches and better assays for the laboratory diagnosis of Lyme disease. All attempts to develop a practical assay for the direct detection of Borrelia burgdorferi in infected patients have failed. Thus far, all proposed alternatives to serology have been unsuccessful, not cost-effective, or are still in early development. Improving serological detection in early disease is the fastest and most effective way to improve patient outcomes in Lyme disease. The antigen targets utilized in current serodiagnostic assays have considerable defects. They often contain conserved epitopes that cross-react with antibodies raised to other antigens, reducing specificity and requiring the use of a two-tier seroassay paradigm that preserves specificity at the cost of poor sensitivity in the detection of early disease. The use of peptides as serodiagnostic targets demonstrate improved efficacy, but the use of one or two peptides containing single epitopes still provides re- duced sensitivity in early disease. Putting the same old antigen targets into new platforms, no matter how in- novative the platform, will not succeed in improving serodiagnostics for Lyme disease. Both the target anti- gens and assays need an innovative approach. By incorporating unique peptides containing linear epitopes highly specific to B. burgdorferi, into a cutting-edge, multiplex, portable paper-based point-of-care diagnostic assay that uses a cost-effective smartphone-based reader, we aim to transform the diagnosis of Lyme disease. An ideal test for Lyme disease could be performed in a single step and yield an answer on the spot to support diagnosis and direct the course of clinical treatment. We have developed an innovative vertical-flow assay which allows for the multiplexed detection of IgM and IgG binding of up to 25 independent antigen targets in a point-of-care setting. The VFA design allows for uniform flow of sample across the target membrane, creating uniform binding conditions and maximizing developed signal. We coupled this design with a cost-effective portable smart-phone based reader allowing for quantitative measure of antibody binding, eliminating subjectiv- ity. By multiplexing peptide antigens each containing 1-2 epitopes unique to B. burgdorferi from multiple differ- ent antigens expressed at different stages during mammalian infection we can generate a single tier, POC as- say that can specifically and sensitively detect patient antibody at all stages of the disease. The assay can be completed in less than 25 min allowing for rapid in office results to support clinical diagnosis, thereby improving patient outcomes.

抽象的 项目摘要显然需要新方法和更好的测定莱姆病实验室诊断。所有试图开发用于直接检测被感染患者Borrelia burgdorferi的实用测定方法的尝试都失败了。到目前为止，所有提议的血清学替代方法都没有成功，没有成本效益，或者仍在早期发展。改善早期疾病的血清学检测是改善莱姆病患者结局的最快，最有效的方法。当前血清诊断测定中使用的抗原靶标具有相当大的缺陷。它们通常包含保守的表位，这些表位与对其他抗原饲养的抗体进行交叉反应，降低特异性并需要使用两层血清测定范式，以在早期疾病检测中以敏感性较差，以保持特异性。将肽用作血清诊断靶标的使用表明功效提高，但使用一个或两个含有单个表位的肽仍然可以在早期疾病中具有敏感性。将相同的旧抗原靶标放入新的平台中，无论该平台如何，都将无法成功改善莱姆病的血清诊断。目标反性和测定都需要一种创新的方法。通过将含有高度特定于B. burgdorferi的线性表位的独特肽结合到使用基于成本效益的智能手机读取器的尖端，多重的，基于纸张的诊断测定器中，我们旨在改变莱姆病的诊断。可以单步进行莱姆病的理想测试，并在现场提供答案，以支持诊断并指导临床治疗过程。我们已经开发了一种创新的垂直流分析，该测定法可以在护理点环境中对IgM和IgG结合多达25个独立抗原靶标的多路复用检测。 VFA设计允许在整个目标膜上均匀的样品流，从而产生均匀的结合条件并最大化开发的信号。我们将这种设计与基于智能手机的可移植智能手机的读取器相结合，允许对抗体结合的定量度量，从而消除了主题。通过将肽抗原多路复用抗原，每种抗原含有1-2个表位的B. burgdorferi所特有的来自多种不同抗原在哺乳动物感染期间在不同阶段表达的抗原所特有的，我们可以产生一个单一的poc，例如，在疾病的所有阶段都可以特异性地检测患者抗体。该测定方法可以在不到25分钟的时间内完成，以使办公室成果快速支持临床诊断，从而改善患者的预后。